a Department of Pediatrics, Henan Xinxiang Central Hospital , Xinxiang , China.
Artif Cells Nanomed Biotechnol. 2019 Dec;47(1):3072-3078. doi: 10.1080/21691401.2019.1637886.
To investigate the potential mechanism of microRNA-34a (miR-34a) on proliferation, migration and invasion of paediatric neuroblastoma cells. The expression of miR-34a and hepatocyte nuclear factor 4α (HNF4α) in paediatric neuroblastoma tissues were detected by RT-q PCR and Western blot, respectively. Cell proliferation, migration, invasion and the expression of matrix metalloproteinase 2 (MMP-2) and matrix metalloproteinase 14 (MMP-14) after transfection of miR-34a mimics or HNF4α siRNA into SH-SY5Y cells were detected by MTT assay, Transwell assay and Western blot assay, respectively. The target relationship between miR-34a and HNF4α was verified by TargetScan online prediction and dual-luciferase assay. Cell proliferation, migration and invasion of SH-SY5Y cells after overexpression of miR-34a and HNF4α were detected. The expression level of miR-34a was decreased ( < .05) while the expression level of HNF4α was increased ( < .05) in paediatric neuroblastoma tissues. Over- expression of Mi-34a or knockdown of HNF4α in SH-SY5Y cells could lead to a decreased of cell proliferation, migration, invasion and the expression of MMP-2 and MMP-14 ( < .05). The results of TargetScan online prediction and dual-luciferase assay indicted that HNF4α was a potential target gene for miR-34a. Overexpression of HNF4α could reverse the inhibition of miR-34a on proliferation, migration and invasion of SH-SY5Y cells. The expression of miR-34a was down-regulated in paediatric neuroblastoma tissues, and overexpression of miR-34a could inhibit proliferation, migration and invasion of SH-SY5Y cells by targeting HNF4α.
为了研究微小 RNA-34a（miR-34a）对小儿神经母细胞瘤细胞增殖、迁移和侵袭的潜在机制。分别采用 RT-qPCR 和 Western blot 检测小儿神经母细胞瘤组织中 miR-34a 和肝细胞核因子 4α（HNF4α）的表达。转染 miR-34a 模拟物或 HNF4α siRNA 后，通过 MTT 测定法、Transwell 测定法和 Western blot 测定法分别检测 SH-SY5Y 细胞的增殖、迁移、侵袭以及基质金属蛋白酶 2（MMP-2）和基质金属蛋白酶 14（MMP-14）的表达。通过 TargetScan 在线预测和双荧光素酶报告基因实验验证 miR-34a 与 HNF4α 的靶关系。检测过表达 miR-34a 和 HNF4α 后 SH-SY5Y 细胞的增殖、迁移和侵袭。小儿神经母细胞瘤组织中 miR-34a 表达水平降低（ < .05），HNF4α 表达水平升高（ < .05）。SH-SY5Y 细胞中 miR-34a 的过表达或 HNF4α 的敲低可导致细胞增殖、迁移、侵袭以及 MMP-2 和 MMP-14 的表达降低（ < .05）。TargetScan 在线预测和双荧光素酶报告基因实验的结果表明，HNF4α 是 miR-34a 的潜在靶基因。过表达 HNF4α 可逆转 miR-34a 对 SH-SY5Y 细胞增殖、迁移和侵袭的抑制作用。miR-34a 在小儿神经母细胞瘤组织中表达下调，过表达 miR-34a 可通过靶向 HNF4α 抑制 SH-SY5Y 细胞的增殖、迁移和侵袭。
